Requirement of replicating RSV, NF-κB induction, and
paracrine/autocrine action of TNF in HBD2 induction. A, total RNA
collected from mock-infected/untreated (UT) and TNF-treated (treated
with 10 ng/ml TNF for 16 h) or RSV-infected (infected with 0.5 m.o.i. virus
for 16 h) A549 cells infected with recombinant adenovirus expressing either
GFP (control) (Ad-GFP) or IκB-super-repressor (Ad-SR)
were subjected to RT-PCR to detect HBD2 and GAPDH (loading control).
B, total RNA collected from untreated (UT)/mock-, TNF (10
ng/ml for 16–24 h)-treated and RSV-infected (infected with 0.5 m.o.i.
virus for 16 and 24 h) A549 cells expressing either GFP or IκB-SR
(SR) were subjected to real time qPCR to detect HBD2 expression.
C, total RNA collected from mock-infected and RSV-infected cells
(infected with either live/non-UV or UV-irradiated RSV) were subjected to
RT-PCR to detect HBD2. HBD2 mRNA expression was also monitored in cells
infected with RSV in the presence of control antibody (control Ab) or
TNF-neutralizing antibody (TNF Ab). D, total RNA collected
from mock, live RSV (non-UV), and UV inactivated (UV)
RSV-infected A549 cells either treated with control antibody (control
Ab) or TNF-neutralizing antibody (TNF Ab) were subjected to qPCR
to detect HBD2 expression. For the qPCR, the transcript levels were normalized
to human keratin 5, and the results are representative of three independent
experiments with similar values.